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LINE-1 Methylation Level and Patient Prognosis in a Database of 208 Hepatocellular Carcinomas

  • Gastrointestinal Oncology
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Abstract

Background

The level of long interspersed nucleotide element-1 (LINE-1) methylation has become regarded as a surrogate marker of global DNA methylation. Previously, we demonstrated that LINE-1 hypomethylation might contribute to the acquisition of aggressive tumor behavior through genomic gains of oncogenes such as cyclin-dependent kinase 6 (CDK6) in esophageal squamous cell carcinoma. However, the relationship between LINE-1 hypomethylation and clinical outcome in hepatocellular carcinoma (HCC) remains unclear.

Methods

LINE-1 methylation level in 208 samples of curatively resected HCCs was measured by pyrosequencing assay, and the prognostic value of LINE-1 methylation level in HCC was examined.

Results

LINE-1 methylation levels in the 208 HCC patients investigated were distributed as follows: mean 64.7; median 64.6; standard deviation (SD) 13.6; range 21.5–99.1; interquartile range 62.9–66.6. Univariate Cox regression analysis revealed a significantly higher cancer recurrence rate in the low-methylation-level group than in the high-methylation-level group (hazard ratio 1.58; 95 % CI 1.05–2.47; p = 0.028). Interestingly, the influence of LINE-1 hypomethylation on patient outcome was modified by hepatitis virus infection (p of interaction = 0.023); LINE-1 hypomethylation was associated with a higher cancer recurrence rate in patients without hepatitis virus infection (log-rank p = 0.0047). CDK6 messenger RNA expression levels were inversely associated with LINE-1 methylation levels (p = 0.0075; R = −0.37).

Conclusions

Genome-wide DNA hypomethylation, as measured by LINE-1 levels, might be associated with poor disease-free survival in HCC patients, suggesting a potential role for LINE-1 methylation level as a biomarker for identifying patients who will experience an unfavorable clinical outcome.

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Acknowledgments

Author contributions: conception and design: Kazuto Harada, Yoshifumi Baba, Toru Beppu, and Hideo Baba; acquisition of data: Kazuto Harada, Yoshifumi Baba, and Toru Beppu; analysis and interpretation of data: Kazuto Harada and Yoshifumi Baba; manuscript writing: Kazuto Harada, Yoshifumi Baba, and Hideo Baba. All authors approved the final manuscript.

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No conflicts of interest exist.

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Authors and Affiliations

  1. Department of Gastroenterological Surgery, Graduate School of Medical Science, Kumamoto University, Kumamoto, Japan

    Kazuto Harada MD, Yoshifumi Baba MD, PhD, Takatsugu Ishimoto MD, PhD, Akira Chikamoto MD, Keisuke Kosumi MD, Hiromitsu Hayashi MD, PhD, Hidetoshi Nitta MD, PhD, Daisuke Hashimoto MD, PhD, Toru Beppu MD, PhD & Hideo Baba MD, PhD

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Correspondence to Hideo Baba MD, PhD.

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10434_2014_4134_MOESM1_ESM.tif

Kaplan–Meier curves for the study group (red) and excluded group (blue). The left and right panels show theoverall survival rate and the disease-free survival rate, respectively (TIFF 768 kb)

10434_2014_4134_MOESM2_ESM.tif

The pyrosequencing assay used to measure the LINE-1 methylation level. The overall LINE-1methylationlevel is the average proportion of C (%) at the 4 CpG sites. The percentages of C at each CpG site afterbisulfite conversion, used to compute the methylation level at each CpG site, are given in blue font. Upperand lower panels show the results for LINE-1 hypermethylated tumor (methylation level, 81.5%) and LINE-1hypomethylated tumor (methylation level, 48.3%), respectively (TIFF 2064 kb)

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Harada, K., Baba, Y., Ishimoto, T. et al. LINE-1 Methylation Level and Patient Prognosis in a Database of 208 Hepatocellular Carcinomas. Ann Surg Oncol 22, 1280–1287 (2015). https://doi.org/10.1245/s10434-014-4134-3

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  • DOI: https://doi.org/10.1245/s10434-014-4134-3

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